The objectives of the proposed research are to prepare highly purified and specific immunological probes to be used in further defining virus gene expression within herpes simplex virus type 1 (HSV-1)- and type 2 (HSV-2)-infected and transformed cells. The nonglycosylated proteins obtained from purified virions and from infected cells will be fractionated and purified by two general approaches. First, HSV-specific polypeptides will be purified by high resolution preparative polyacrylamide gel electrophoresis containing sodium dodecyl sulfate. These highly purified polypeptides will be further purified and analyzed by two-dimensional gel electrophoresis. Second, HSV-specific native proteins will be purified utilizing several techniques which include hydroxylapatite chromatography, isoelectric focusing and affinity chromatography. Monospecific antisera to the purified polypeptides and native proteins will be produced in rabbits. These antisera will be used in studies to further characterize the proteins synthesized in the infected cell incorporating the use of immunofluorescence, immunoprecipitation, radioimmunoassay, and immunoperoxidase staining. In addition, these sera will be used extensively to define the expression of HSV specific genes within the transformed cell. It is postulated that such highly specific immunological probes will eventually be of significant value in further defining the role certain HSV proteins may play in the latently infected cell as well as in the cancer cell.